In Vitro Spectroscopy-Based Profiling of Urothelial Carcinoma: A Fourier Transform Infrared and Raman Imaging Study

Abstract

Markers of bladder cancer cells remain elusive, which is a major cause of the low recognition of this malignant neoplasm and its recurrence. This implies an urgent need for additional diagnostic tools which are based on the identification of the chemism of bladder cancer. In this study, we employed label-free techniques of molecular imaging—Fourier Transform Infrared and Raman spectroscopic imaging—to investigate bladder cancer cell lines of various invasiveness (T24a, T24p, HT-1376, and J82). The urothelial HCV-29 cell line was the healthy control. Specific biomolecules discriminated spatial distribution of the nucleus and cytoplasm and indicated the presence of lipid bodies and graininess in some cell lines. The most prominent discriminators are the total content of lipids and sugar moieties as well as the presence of glycogen and other carbohydrates, un/saturated lipids, cytochromes, and a level of S-S bridges in proteins. The combination of the obtained hyperspectral database and chemometric methods showed a clear differentiation of each cell line at the level of the nuclei and cytoplasm and pointed out spectral signals which differentiated bladder cancer cells. Registered spectral markers correlated with biochemical composition changes can be associated with pathogenesis and potentially used for the diagnosis of bladder cancer and response to experimental therapies.

FTIR Spectroscopic Imaging Supports Urine Cytology for Classification of Low- and High-Grade Bladder Carcinoma

“…And all of these low-glycogen cells exhibited high nuclear-cytoplasmic ratios. These observations were congruent with our previous spectroscopic studies on urine sediments and the reference cell cultures of urothelial and BC cells [11,14]. Spectral differences between the N and BC groups were seen the shapes of the glycogen bands, e.g., a sharp 1024 cm −1 band was present in N while an additional 1054 cm −1 band was found in both BC groups.…”

“…These similarities resulted from the biological nature of proliferating cells, i.e., cells that proliferate quickly, have a low cytoplasm to nuclear ratio and have cytoplasm can contain spare substances. These differences between nuclei and cytoplasm were discussed in a report showing the IR features of urinary cell cultures detected by high-definition FTIR imaging [11]. Here, scanning of cytological samples with a focal plane array of a 5.5 × 5.5 µm pixel size did not allow for the observation of carbohydrates in the cell compartments.…”

“…However, no patients' cytological cells have yet been imaged with detection through FTIR and Raman spectra. The only urothelial cell imaging to date was performed on cell cultures [11,12]. A single-point microscopic examination of urothelial cells in urine from healthy volunteers showed the differentiation of three cell types only [10].…”

Raman spectroscopy biochemical characterisation of bladder cancer cisplatin resistance regulated by FDFT1: a review

“…Additional file 1: Tables S1 and S3 clearly show that researchers verified the application of Raman spectroscopy on different biological sources in diagnosing bladder cancer. With respect to the specification mentioned, Kujdowicz et al employed cultured cell lines of healthy bladder, low-grade and high-grade bladder cancer in their studies [79].…”

FT-IR Spectral Signature of Sensitive and Multidrug-Resistant Osteosarcoma Cell-Derived Extracellular Nanovesicles

“…Fourier-transform infrared spectroscopy (FT-IR) has been fruitfully applied to describe structural characterization of proteins and protein–membrane interactions in biological matrices [ 11 ]. Malignant cell characteristics and cell differentiation show specific vibrational molecular signatures, thus indicating a modified chemical composition associated with biomolecules such as proteins, nucleic acids, lipids, and carbohydrates [ 12 , 13 , 14 ]. Vibrational spectroscopy is simple, label-free, sensitive, and requires minimal sample preparation.…”